title: Recombinant production of TEV cleaved human parathyroid hormone.
abstract: The parathyroid hormone, PTH, is responsible for calcium and phosphate ion homeostasis in the body. The first 34 amino acids of the peptide maintain the biological activity of the hormone and is currently marketed for calcium imbalance disorders. Although several methods for the production of recombinant PTH(1-34) have been reported, most involve the use of cleavage conditions that result in a modified peptide or unfavorable side products. Herein, we detail the recombinant production of (15) N-enriched human parathyroid hormone, (15) N PTH(1-34), generated via a plasmid vector that gives reasonable yield, low-cost protease cleavage (leaving the native N-terminal serine in its amino form), and purification by affinity and size exclusion chromatography. We characterize the product by multidimensional, heteronuclear NMR, circular dichroism, and LC/MS.
authors.affiliation: Department of Chemistry, Dartmouth College, Hanover, New Hampshire 03755, USA.
authors.forename: Christopher O
authors.initials: CO
authors.lastname: Audu
authors.forename: Jared C
authors.initials: JC
authors.lastname: Cochran
authors.forename: Maria
authors.initials: M
authors.lastname: Pellegrini
authors.forename: Dale F
authors.initials: DF
authors.lastname: Mierke
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002846:Chromatography, Affinity; D010450:Endopeptidases / Q000737:chemistry*; D004926:Escherichia coli; D006801:Humans; D008969:Molecular Sequence Data; D010281:Parathyroid Hormone / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D017433:Protein Structure, Secondary; D059748:Proteolysis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D010281:Parathyroid Hormone; D011993:Recombinant Fusion Proteins; D010450:Endopeptidases; C099488:TEV protease
keywords: LC/MS; NMR; TEV protease; circular dichroism; parathyroid hormone; recombinant peptide production